Addgene - pRK5 BARK1 minigene Plasmid Data

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Plasmid 14695: pRK5 BARK1 minigene

Gene/insert name:		BARK1 minigene
Alternative names:		beta ARK1
		beta adrenergic receptor kinase
		ADRBK1
Insert size (bp):		Unknown
Gene/insert aliases:		ADRBK1, GRK2
Species of gene(s):		B. taurus (bovine)
Relevant mutations/deletions:		BARK1 amino acids 495-689
Fusion proteins or tags:		3'UTR of human b-globin
Terminal:		C terminal on insert
Vector backbone:		pRK5 (Search Vector Database)
Type of vector:		Mammalian expression
Backbone size (bp):		4700
Cloning site 5':		EcoRI
Site destroyed during cloning:		No
Cloning site 3':		XbaI
Site destroyed during cloning:		No
5' Sequencing primer:		Sp6 (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		High Copy
Grow in standard E. coli @ 37C:		Yes
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		DH5a
Principal Investigator:		Robert J. Lefkowitz
Terms and Licenses:		MTA

Comments: The cDNA encoding bovine BARK1 residues Gly495 to Leu689 was amplified using PCR to construct a minigene casette with newly engineered 5' and 3' ends (see Author's map). The 5' end contained an EcoRI site for subcloning, followed by the kozak sequence 5'-GCCGCCACCATGG-3'. A bclI site was inserted at the 3' end immediately following the stop codon. The final BARK1 minigene construct contains the 3'UTR of the human beta-globin gene and a polyadenylation signal.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

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Article: Cellular expression of the carboxyl terminus of a G protein-coupled receptor kinase attenuates G beta gamma-mediated signaling. Koch WJ et al. (J Biol Chem. 1994 Feb 25. 269(8):6193-7. Pubmed)

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